Journal: Nature protocols

Article Title: Lentiviral transduction of mammalian cells for fast, scalable and high-level production of soluble and membrane proteins

doi: 10.1038/s41596-018-0075-9

Figure Lengend Snippet: List of transfer plasmids.

Article Snippet: Proviral DNA sizes are noted in brackets. (D) Comparative size-exclusion chromatograms of secreted NET1 ΔNTR and NEO1 FN456 , showing improved secreted protein yields from lentiviral transduction (blue curves) as compared to transient transfection (red curves) of adherent HEK293T cells. table ft1 table-wrap mode="anchored" t5 caption a7 Plasmid no. pHR-CMV-TetO 2 Addgene Plasmid # Purpose 1 3C-Twin-Strep 113883 Protein (co-)expression, FACS 2 3C-Twin-Strep_IRES-EmGFP 113884 3 3C-Twin-Strep_IRES-mRuby2 113885 4 3C-Twin-Strep_IRES-mTurquoise2 113886 5 3C-Avi-His6 113887 Protein (co-)expression, FACS, in vivo biotinylation 6 3C-Avi-His6_IRES-EmGFP 113888 7 3C-Avi-His6_IRES-mRuby2 113889 8 3C-Avi-His6_IRES-mTurquoise2 113890 9 3C-mVenus-Twin-Strep 113891 Membrane protein expression and FSEC-TS screening, FACS 10 EmGFP 113892 Transduction & FACS compensation controls 11 mVenus 113893 12 mRuby2 113894 13 mTurquoise2 113895 14 HA-BirA 113896 HA-tagged cytosolic and ER-resident biotin ligase for in vivo biotinylation 15 HA-BirA-ER 113897 16 3C-Avi-His6_IRES-HA-BirA-ER 113898 Plasmid no. pHR-CMV Addgene Plasmid # Purpose 17 TetR-HA-NLS-P2A-BSD-Myc 113899 Generation of inducible cell lines Plasmid no. pHR-SFFV Addgene Plasmid # Purpose 18 3C-Twin-Strep 113900 Alternative for CMV-TetO 2 Plasmid no. pHR-CAG Addgene Plasmid # Purpose 19 3C-Twin-Strep 113901 Alternative for CMV-TetO 2 Open in a separate window caption a8 List of transfer plasmids.

Techniques: Plasmid Preparation, In Vivo, Expressing, Transduction